WO2025003171A1

WO2025003171A1 - Device for percutaneous cryolipolysis, kit for percutaneous cryolipolysis and use of such a device or kit for percutaneous cryolipolysis

Info

Publication number: WO2025003171A1
Application number: PCT/EP2024/067868
Authority: WO
Inventors: Carolin WINTER
Original assignee: Merz Aesthetics GmbH
Current assignee: Merz Aesthetics GmbH
Priority date: 2023-06-30
Filing date: 2024-06-25
Publication date: 2025-01-02
Anticipated expiration: 2025-12-30
Also published as: AU2024306077A1

Abstract

The present invention relates to a device for percutaneous cryolipolysis, to a kit for percutaneous cryolipolysis and to the use of such a device or kit for percutaneous cryolipolysis, wherein the device comprises a main body, to which a cryo fluid source is couplable, and a cryo needle assembly (11), being coupled or couplable to the main body, wherein the cryo needle assembly (11) comprises at least one cryo needle (12). The at least one cryo needle (12) is configured to penetrate the skin (D1, D2) of a human or an animal at least partially. The device is configured to cause a cooling of at least one cryo needle (12) at least in a cryo needle cooling area (P1), when a cryo fluid source and the cryo needle assembly (11) are coupled to the main body, by using cryo fluid (G) from the cryo fluid source, and to form a subcutaneous (F) tissue cooling zone (CZ) adjacent to the cryo needle cooling area (P1) of the cooled cryo needle (12), when the cooled cryo needle (12) is penetrating the skin (D1, D2) of a human or an animal at least partially, wherein the device is configured to cool subcutaneous (F) fat cells (A, A1, A2) located in the tissue cooling zone (CZ) such that at least for some of the fat cells (A, A1, A2) located in the tissue cooling zone (CZ) apoptosis (A1) and/or necrosis (A2) is initiated.

Description

DEVICE FOR PERCUTANEOUS CRYOLIPOLYSIS, KIT FOR PERCUTANEOUS CRYOLIPOLYSIS AND USE OF SUCH A DEVICE OR KIT FOR PERCUTANEOUS CRYOLIPOLYSIS

FIELD OF THE INVENTION

The present invention relates to a device for percutaneous cryolipolysis, wherein the device comprises a main body, to which a cryo fluid source is couplable, and a cryo needle assembly, being coupled or couplable to the main body, wherein the cryo needle assembly comprises at least one cryo needle. At least one cryo needle is configured to penetrate the skin of a human or an animal at least partially. The device is configured to cause a cooling of at least one cryo needle at least in a cryo needle cooling area, when a cryo fluid source and the cryo needle assembly are coupled to the main body, by using cryo fluid from the cryo fluid source, and the device is configured to form a subcutaneous tissue cooling zone adjacent to the cryo needle cooling area of the cooled cryo needle, when the cooled cryo needle is penetrating the skin of a human or an animal at least partially.

The present invention further relates to a kit for percutaneous cryolipolysis, comprising a device for percutaneous cryolipolysis and a cryo fluid source.

In addition, the present invention relates to the use of a device for percutaneous cryolipolysis or a kit for percutaneous cryolipolysis for cosmetic, aesthetic or therapeutic application, particularly for percutaneous cryolipolysis for cosmetic, aesthetic or therapeutic purpose.

BACKGROUND OF THE INVENTION

Sometimes treatment of body tissue, in particular destruction and/or removal of body tissue like tumor tissue or fat tissue, is wanted or necessary, from a health perspective or for other reasons, for example for cosmetic and/or aesthetic reasons.

One established therapy for the treatment of tumor tissue, in particular for destruction and/or removal of tumor tissue, for example, is so-called “cryosurgery”, in particular cryoablation of tumor tissue, which is based on application of extreme cold to the tumor tissue to be treated. Several methods and devices are known therefore.

WO 2019/213205 A1 , for example, discloses a handheld cryoprobe for use in invasive, percutaneous cryotherapy of tumorous masses. The handheld cryoprobe includes a probe attached to a CO2 gas dispensing backend. The probe has specifically optimized parameters designed for use with CO2 gas and is made out of a partially hollowed and threaded aluminium rod providing maximum heat exchange. The system backend regulates flow of compressed CO2 gas while throttling and cooling the gas coolant to the cytotoxically low temperatures necessary for targeted tumour cell death. Additionally, the incoming initial stream of CO2 gas is throttled by the Joule-Thomson nozzle on the backend. The low temperature exhaust gas is then used to pre-cool all subsequent incoming gas, resulting in an even lower temperature at the probe tip, which provides a positive feedback loop, continually decreasing the gas's temperature. The temperature drop is caused by the Joule-Thomson effect.

From US 2021/244457 A1 , an apparatus for invasive treatment of a breast, a prostate or a kidney tumour is known, consisting of a probe, containing a lumen and having a distal end configured to contact tissue of a living subject. A temperature sensor is located at the distal end, and a pump, having a pump motor, is coupled to deliver a cryogenic fluid through the lumen to the distal end of the probe and to receive the cryogenic fluid returning from the probe. There is a separator, coupled to separate the returning cryogenic fluid into a returning cryogenic liquid and a returning cryogenic gas, and a flow meter, coupled to measure a rate of flow of the returning cryogenic gas. A processor is configured to control a rate of pumping of the pump motor in response to a temperature measured by the temperature sensor and the rate of flow of the returning cryogenic gas.

The company “IceCure” provides with “ProSense™” a cryoablation device for treatment of tumorous tissue of several cancer types, in particular for treatment of breast cancer and prostate, lung and bone cancer, which uses liquid nitrogen for cooling, see for example https://icecure-medical.com/ and https://youtu.be/TfhQJ3SN6wQ, last accessed on June 14, 2023.

Boston Scientific also provides cryoablation devices for invasive treatment of tumour cells by applying extreme cold, wherein the tumour cells are cold down to -20°C to -40°C, see for example https://www.bostonscientific.com/en-

US/products/cryoablation/icefx.html#, last accessed on June 14, 2023.

Fat cells, which are different from tumour cells and have different properties and show different reactions on treatment, are often removed by liposuction, wherein fat cells are separated from the surrounding tissue and are suctioned off. However, liposuction is an invasive method and has a plurality of undesirable side effects.

For improved liposuction, WO 98/41157 suggests combining cryosurgery and liposuction, wherein first by cryosurgery fatty tissue to be removed is destroyed by controlled freezing of the tissue. This facilitates removal of the fatty tissue. Subsequently, by liposuction the destroyed fatty tissue is removed by aspiration.

Other methods to reduce fatty tissue, in particular its volume, are based on the destruction of the fat cells by lipolysis, wherein different methods for triggering/initiation of lipolysis are known.

Lipolysis of fat cells may for example be triggered by injection of a defined composition which causes cell death of the fat cells into the fatty tissue.

Such a composition is for example “Kybella®” by Allergan Pharmaceuticals International Limited, which comprises desoxycholic acid as initiator for lipolysis. However, undesirable side effects may occur.

Another method which is based on injection of a composition to trigger lipolysis is described in Ni P, Farinelli WA, Cheng LL, Farrar CT, Motamarry A, Moradi Tuchayi S, Wang Y, Anderson RR, Garibyan L. Total ice content and lipid saturation determine adipose tissue cryolipolysis by injection of ice-slurry. Lasers Surg Med. 2023 Jan;55(1):116-125. doi: 10.1002/lsm.23557. Epub 2022 May 21. PMID: 35598082;

PMCID: PMC9676409, available under https://pubmed.ncbi.nlm.nih.gov/35598082/, last accessed on June 14, 2023. This method is based on injection of a defined “ice-slurry”.

One significant disadvantage of methods for lipolysis based on injection of a composition into the fatty tissue is that once the composition is injected, the reaction of the tissue can no longer be influenced and the process can no longer be controlled. In addition, it is known to destroy fat cells by use of non-invasive cryolipolysis, wherein lipolysis is caused by freezing the fat cells from outside, wherein the cold is applied via the skin. However, this method entails risks such as superficial cold burns of the skin. Devices for non-invasive cryolipolysis treatment of fat tissue are for example provided by Allergan Aesthetics respectively ZELTIQ™ Aesthetics, Inc, wherein these devices are promoted as “Coolsculpting”-devices, see for example https://www.coolsculpting.com/, last accessed on June 14, 2023. With these devices, the body fat tissue is frozen by cold applied via the skin. One unwanted side effect known for this kind of treatment is paradoxical adipose hyperplasia.

Furthermore, using cold for treatment is known for pain therapy. Under iovera®, for example, a device for pain therapy using extreme cold to stop nerves from sending pain signals to the brain is promoted, see for example https://www.iovera.com/, last accessed on June 14, 2023.

OBJECTIVES OF THE INVENTION

In view of the above, one objective of the present invention is the provision of a device, which enables improved treatment of fatty tissue, in particular improved destruction and/or improved removal of fat cells, preferably with reduced or better controllable side effects.

Another objective of the present invention is the provision of a kit, which enables improved treatment of fatty tissue, in particular improved destruction and/or improved removal of fat cells, preferably with reduced or better controllable side effects.

A further objective of the present invention is the provision of a method for use, by which improved treatment of fatty tissue, in particular improved destruction and/or improved removal of fat cells can be achieved, preferably with reduced or better controllable side effects.

SUMMARY OF THE INVENTION

These objects of the present invention are achieved by providing a device, a kit and method for use as defined in the independent claims. Possible and in particular preferred embodiments of the objects of the present invention are set forth in the dependent claims. Further embodiments and other objects, advantages and features of the present invention will become apparent from the following detailed description of the invention, the illustration of particular embodiments in the accompanying figures and the detailed description of the embodiments. By express reference, the wording of the original claims is herewith incorporated to the description.

Some objects of the present invention are in particular achieved by a device for percutaneous cryolipolysis, wherein the device comprises a main body, to which a cryo fluid source is couplable, and a cryo needle assembly, being coupled or couplable to the main body, wherein the cryo needle assembly comprises at least one cryo needle. At least one cryo needle of said device is configured to penetrate the skin of a human or an animal at least partially. Said device is configured to cause a cooling of at least one cryo needle at least in a cryo needle cooling area, when a cryo fluid source and the cryo needle assembly are coupled to the main body, by using cryo fluid from the cryo fluid source, and the device is configured to form a subcutaneous tissue cooling zone adjacent to the cryo needle cooling area of the cooled cryo needle, when the cooled cryo needle is penetrating the skin of a human or an animal at least partially. Said device is further configured to cool subcutaneous fat cells located in the tissue cooling zone such that at least for some of the fat cells located in the tissue cooling zone apoptosis and/or necrosis is initiated.

Some objects of the present invention are in particular achieved by a kit comprising such a device and a cryo fluid source.

Some objects of the present invention are in particular achieved by use of such a device and/or such a kit.

DETAILED DESCRIPTION OF THE INVENTION

A device for percutaneous cryolipolysis according to the present invention may comprise a main body, to which a cryo fluid source may be couplable, and a cryo needle assembly, being coupled or couplable to the main body, wherein the cryo needle assembly may comprise at least one cryo needle. The at least one cryo needle may be configured to penetrate the skin of a human or an animal at least partially. The device may be configured to cause a cooling of at least one cryo needle at least in a cryo needle cooling area, when a cryo fluid source and the cryo needle assembly are coupled to the main body, by using cryo fluid from the cryo fluid source, and to form a subcutaneous tissue cooling zone adjacent to the cryo needle cooling area of the cooled cryo needle, when the cooled cryo needle is penetrating the skin of a human or an animal at least partially. The device may further be configured to cool subcutaneous fat cells located in the tissue cooling zone such that at least for some of the fat cells located in the tissue cooling zone apoptosis and/or necrosis is initiated.

With a device according to the present invention, it seems that at least in some cases and at least in some regions of a human body subcutaneous fat tissue can be destroyed and removed precisely at lower risk, with increased safety and reduced side effects compared to other methods like liposuction, “slurry-injection”, “Kybella® or “CoolSculpting®”. A device according to the invention appears to allow precise lipolysis of subcutaneous fat cells in a wide variety of indications, such as for example lipoedema and/or lipoma, and/or other reasons.

A device according to the present invention seems in particular to allow a targeted utilization, in particular an advantageous utilization, of different cell death mechanisms, depending on the respective needs and/or use cases. In particular, it seems possible with a device according to the invention to initiate a desired cell death mechanism for different cell layers in each case. By a device according to the invention, lipoma may for example be treated by initiating apoptosis for the inner core-structure, and initiating necrosis for the outer shell-structure. First necrosis and then apoptosis is also possible, or a mixture of apoptosis and necrosis. This could be done in one treatment session or in more sessions.

The term “cryolipolysis” in the meaning of the disclosure herein means causing cell death of human or animal fat cells by applying cold to the cell. The term “cryolipolysis” means in particular causing cell death of the so-called “white” fat cells, which are also often named as “adipocytes” and/or “mature adipocytes” and which are configured to store high amounts of fat in the human or animal body.

The term “percutaneous” in the meaning of the disclosure herein means “by punctuation through the skin” according to its common and established medical definition.

The term “subcutaneous” in the in the meaning of the disclosure herein means “means under or beneath the skin” according to its common and established medical definition.

The main body may comprise a housing and may be designed ergonomically for good handling by a practitioner. The main body may in particular comprise a gripping zone. The main body may also comprise one or more control buttons for control of the device, in particular a power button and/or one or more switches to switch on and/or off the cooling, wherein in particular the flow of the cryo fluid may be controlled via operating elements, which may be arranged or integrated in the main body. The main body may in particular be made of a lightweight material or comprises at least partly a lightweight material to allow precise handling for precise cryo application. In addition or alternatively, one or more control buttons and/or operating elements may be arranged in a separate terminal, which can be coupled directly or indirectly with the main body as it is known from dental devices.

A “cryo fluid source” in the meaning of the present disclosure is a source of fluid, wherein the term “fluid” encompasses gas and liquid mediums, wherein the fluid, which is provided by the source, may cause cooling of a cryo needle, in particular of an outer surface of a cryo needle, when flowing through the cryo needle. A cryo fluid source may, for example be, but is not limited to, a container or a cartridge filled with liquid nitrogen (N2), carbon dioxide (CO2) or nitrous oxide (N2O; laughing gas) or any other fluid which is suitable as cryo fluid. In principle, all fluids are suitable for use as a cryo fluid, by which the so called “Joule-Thomson-Effect” can be caused in an appropriate manner to achieve the desired cooling effect. Hence, also other fluids which are well-known from other use cases may be used as a cryo fluid in principle. Other fluids, which might in principle be used are, for example, cooling fluids known from the automotive industry or from air conditioning as in particular R134a, R123yf, R1234ze(E), R404A/ R507, R407A / R407F, R407C, R410A, R32, R23/ R508A/ R508B, R600a, R290 / R1270 or R717. However, when a non-medical cryo fluid is used, additional measures might be required, to ensure safety.

In some embodiments the cryo fluid source may be integrated into the device. In other embodiments, the cryo fluid source may be not integrated into the device and be configured to be located separately, in a separate terminal for example, and be coupled or couplable by a line, a hose or a tube with the device, in particular with the cryo needle, in particular via the main body.

The cryo fluid source may be configured for stationary arrangement and may in particular be a stationary fluid storage, which may, for example, comprise a volume of up to 11, 51, 101, 151 or up to 201 of cryo fluid. However, also other sizes are possible.

In another embodiment, the cryo fluid source may be configured for mobile use. In one embodiment, the cryo fluid source may be, for example, a cryo fluid cartridge. Such a cryo fluid cartridge may in particular have a volume of at least, for example, 0.1 ml, 0.2 ml, 0.3 ml, 0.4 ml, 0.5 ml, 0.6 ml, 0.7 ml or 0.9 ml up to 11, 1.51 or 21 for example. However, also other sizes are possible.

The cryo fluid source may be coupled to the main body as it is for example known from cryosurgery devices for tumor treatment.

A ’’cryo needle assembly” in the meaning of the present disclosure is an assembly comprising a cryo needle and one or more other parts, wherein a cryo needle assembly in particular comprises at least a needle hub for mounting the needle assembly to the main body in addition to the cryo needle. The cryo needle assembly may, for example, comprise in addition an end stop to limit penetration depth of the needle when piercing the skin and placing the needle with the cooling area at the desired location in the subcutaneous fat tissue.

The term “needle” in the present disclosure” comprises needles and cannulas as well, in particular sharp needles configured for piercing the skin, and as well blunt cannulas, which may require piercing of the skin by another means first.

A cryo needle of a device according to the present invention preferably comprises at least one inner lumen through which cryo fluid can flow from an inlet in direction of the needle cooling area, in particular towards the needle tip, wherein by flowing of a cryo fluid through said lumen, a cooling of the outer needle surface may be caused according to the so-called and well-known “Joule-Thomson-Effect” in the needle cooling area.

In a preferred embodiment, the cryo needle is designed such that no cryo fluid is directed into the surrounding tissue. Therefore, in a preferred embodiment, the cryo fluid flow may preferably at least partially be returned.

The cryo needle assembly may in general be designed and configured similar to a cryo needle as known, for example from cryosurgery and cryotherapy of tumor treatment. However, a device according to the present disclosure is able to generate a very specific cooling of subcutaneous fat tissue. In particular, the ability of generating a specific and advantageous tissue cooling zone for lipolysis of subcutaneous fat cells differs.

In the context of this application, the term “fat tissue” in particular means a type of tissue which can be found in humans and animals capable of storing energy in the form of fat, wherein “fat tissue” in particular refers to a tissue comprising adipocytes (specific cells capable of storing energy in the form of fat) and/or preadipocytes (cells which have the potential to differentiate into (mature) adipocytes.

In one embodiment, the cryo fluid may at least partially be recycled and turned back into a cryo fluid circulation. Thereby, the consumption of cryo fluid for cryolipolysis can be reduced significantly. In some cases, a reprocessing of the recycled cryo fluid might be necessary, for example a filtration and/or compression. In one embodiment, the device may comprise additional means for reprocessing of recycled cryo fluid and be configured to circulate cryo fluid at least partially.

The cold outer surface of the needle cools the surrounding tissue, especially the tissue adjacent to the outer needle surface. Depending on the amount of generated cooling energy, the temperature of the outer needle surface and size and shape of the needle cooling area, also tissue is cold that is not directly adjacent to the needle, i.e., that is not in direct contact with the needle. In particular, a in the following so-called “tissue cooling zone” is generated.

Preferably, the needle cooling zone is located somewhere along the needle between the needle tip and the connection to the main body.

If the cryo needle is placed and penetrates the skin such that the needle cooling area is at least partially surrounded by subcutaneous tissue, a subcutaneous tissue cooling zone will be generated. Is the surrounding tissue, subcutaneous fat tissue, a subcutaneous fat tissue cooling zone will be generated.

A “subcutaneous tissue cooling zone” in the meaning of this disclosure is a zone located under the skin in human or animal tissue, in particular in subcutaneous fat tissue, in which everywhere within this zone during cooling the temperature of the tissue cells is at least temporarily lower than a defined temperature, wherein the isothermal of said defined temperature defines the boarders of the cooling zone to the surrounding subcutaneous tissue. The defined temperature may in particular be lower than +15°C, +14°C, +13°C, +12°C, +11 °C, +10°C, +9°C, +8°C, +7°C, +6°C, +5°C, +4°C, +3°C, +2°C, +1 ° or 0°C, but not below -50°C, -40°C, -39°C, -38°C, -37°C, -36°C, -35°C, -34°C, -33°C, -32°C, -31°C,-

30°C, -29°C, -28°C, -27°C, -26°C, -25°C, -24°C, -23°C, -22°C, -21 °C, -20°C, -19°C, -18°C,

-17°C, -16°C, -15°C, -14°C, -13°C, -12°C, -11 °C, -10°C, -9°C, -8°C, -7°C, -6°C, -5°C, - 4°C, -3°C, -2°C or -1°C.

In a preferred embodiment, the device may be configured to generate and/or form a tissue cooling zone with a defined size and/or shape. The shape of the subcutaneous tissue cooling zone, in particular within subcutaneous tissue, which can be generated by the device, may in particular at least be partly or completely ellipsoidal, ball-shaped, oval, of a toroid shape, of a conical shape or of a combination thereof. In one embodiment the device may be configured to establish a tissue cooling zone having a shape which is adapted to the body region to be treated by cryolipolysis. This might be achieved by a special cryo needle design, in particular by a specific design of the outer surface of the cryo needle, by a specific design of the inner lumen through which the cryo fluid flows, by an appropriate cryo fluid flow and/or by an appropriate cryo fluid temperature and/or by appropriate insulating properties of the needle, in particular of the needle wall.

The term “apoptosis” has to be understood according to its usual understanding in the medical context and refers to a first mechanism of cell death (controlled cell death). If a cell, in particular a fat cell, had died through apoptosis may, for example, be determined with the help of one or more certain well-known markers.

The term “necrosis” has to be understood according to its usual understanding in the medical context and refers to a second mechanism of cell death (un-controlled cell death). If a cell, in particular a fat cell, had died through necrosis may, for example, also be determined with the help of one or more certain well-known markers.

In one possible embodiment, the device may be configured to cool the at least one cryo needle in the cryo needle cooling area such that for at least 1 Vol.-%, 2 Vol.-%, 3 Vol.-%, 4 VoL-%, 5 Vol.-%, 10 Vol.-%, 15 VoL-%, 20 VoL-%, 25 VoL-%,30 VoL-%, 35 Vol.-%, 40 VoL-%, 45 Vol.-%, 50 Vol.-%, 55 Vol.-%, 60 Vol.-%, 65 VoL-%, 70 VoL-%, 75 Vol.-%, 80 Vol.-%, 85 Vol.-%, 90 Vol.-%, 95 Vol.-% or up to 99 Vol.-% of the fat cells located in the tissue cooling zone apoptosis is initiated.

The device may in one embodiment in particular be configured such that at least for some of the fat cells necrosis is avoided, in particular for the fat cells for which apoptosis is initiated.

In one possible embodiment, in particular in addition or alternatively, the device may be configured to cool the at least one cryo needle in the cryo needle cooling area such that for at least 1 Vol.-%, 2 Vol.-%, 3 Vol.-%, 4 Vol.-%, 5 VoL-%, 10 Vol.-%, 15 Vol.-%, 20 Vol.- %, 25 Vol.-%,30 Vol.-%, 35 Vol.-%, 40 Vol.-%, 45 Vol.-%, 50 Vol.-%, 55 Vol.-%, 60 Vol.- %, 65 Vol.-%, 70 Vol.-%, 75 Vol.-%, 80 Vol.-%, 85 Vol.-%, 90 Vol.-%, 95 Vol.-% or up to 99 Vol.-% of the fat cells located in the tissue cooling zone necrosis is initiated.

The device may in one embodiment in particular be configured such that at least for some of the fat cells apoptosis is avoided, in particular for the fat cells for which necrosis is initiated.

Different mechanisms of cell death may result in different cryolipolysis results, in particular in different long-term results. Side effects may be influenced by the type of cell death, which occurs. The kind of cell death may in particular influence further mechanisms in a human and/or animal body, for example processes for break down and/or removal of the dead fat cells. However, the cryolipolysis results seem not only depend on the kind of cell death, but rather also on several further parameters, as for example the tissue constitution in general, cooling process parameters, in particular cooling temperature and temperature gradient. With a device according to the invention, which allows to control the kind and extent of fat cell death occurring, unwanted side effects can be reduced to a minimum.

In one embodiment, the device may in particular be configured to cause a defined cell death - apoptosis and/or necrosis - in particular with a defined ratio, preferably with a predetermined, wanted ratio between a volume of fat cells died by apoptosis and a volume of fat cells died by necrosis. In some cases apoptosis might be preferred. In other cases necrosis. With a device, which is configured such that the kind of cell death caused by cooling is adjustable at least within defined boarders, a wide spectrum of possible cryolipolysis treatments of different fat cells at different tissue locations can be covered.

In one possible embodiment, the device may be configured to cool the at least one cryo needle in the cryo needle cooling area such that for some fat cells located in the tissue cooling zone apoptosis is initiated and for some fat cells located in the tissue cooling zone necrosis is initiated, wherein in particular the ratio of the volume of fat cells located in the tissue cooling zone for which apoptosis is initiated to the volume of fat cells located in the tissue cooling zone for which necrosis is initiated is in a range from 0.0, 0.1 , 0.2, 0.3, 0.4, 0.5, 0.6, 0.7. 0.8 or 0.9 to 1.0, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2 or 0.1. The ratio may differ for different treatment locations.

The device may in particular be configured to achieve, in particular with one cooling cycle, a ratio between the volume of dead fat cells within the cooling zone for which apoptosis was initiated, and therefore the fat cells in particular died due to apoptosis, to the volume of dead fat cells within the tissue cooling zone, for which necrosis was initiated, and therefore the fat cells in particular died due to necrosis, of 1 :99, 2:98, 3:97, 4:96, 5:95, 6:94, 7:93, 8:92, 9:91 , 10:90, 11 :89, 12:88, 13:87, 14:86, 15:85, 16:84, 17:83, 18:82, 19:81 , 20:80, 21 :79, 22:78, 23:77, 24:76, 25:75, 26:74, 27:73, 28:72, 29:71 , 30:70, 31 :69, 32:68,

33:67, 34:66, 35:65, 36:64, 37:63, 38:62, 39:61 , 40:60, 41 :59, 42:58, 43:57, 44:56, 45:55,

46:54, 47:53, 48:52, 49:51 , 50:50, 51 :49, 52:48, 53:47, 54:46, 55:45, 56:44, 57:43, 58:42, 59:41 , 60:40, 61 :39, 62:38, 63:37, 64:36, 65:35, 66:34, 67:33, 68:32, 69:31 , 70:30, 71 :29. 72:28, 73:27, 74:26, 75:25, 76:24, 77:23. 78:22, 79:21 , 80:20, 81 :19, 82:18, 83:17, 84:16, 85:15, 86:14, 87:13, 88:12, 89:11 , 90:10, 91 :9, 92:8, 93:7, 94:6, 95:5, 96:4, 97:3, 98:2 or 99:1.

In one embodiment, in particular in a preferred embodiment, the device may be configured such that a target ratio of volume of fat cells within the cooling zone for which apoptosis should be initiated to a volume of fat cells within the cooling zone for which necrosis should be initiated, can be adjusted or set.

In one embodiment, in particular in a preferred embodiment, the device may be configured such that size and/or shape of the cooling zone may be adapted.

One important parameter, which seems to influence the kind of cell death which is initiated (apoptosis and/or necrosis) significantly is the cooling temperature.

In one possible embodiment, the device may be configured to cool the at least one cryo needle in the cryo needle cooling area such that in the formed tissue cooling zone the subcutaneous tissue is cold down at least to +15°C, +14°C, +13°C, +12°C, +11 °C, +10°C, +9°C, +8°C, +7°C, +6°C, +5°C, +4°C, +3°C, +2°C, +1° or 0°C, but not cold down below -50°C, -40°C, -39°C, -38°C, -37°C, -36°C, -35°C, -34°C, -33°C, -32°C, -31°C,-

30°C, -29°C, -28°C, -27°C, -26°C, -25°C, -24°C, -23°C, -22°C, -21 °C, -20°C, -19°C, -

18°C, -17°C, -16°C, -15°C, -14°C, -13°C, -12°C, -11 °C, -10°C, -9°C, -8°C, -7°C, -6°C, -

5°C, -4°C, -3°C, -2°C or -1°C, wherein the subcutaneous tissue in the tissue cooling zone is preferably cold down to a tissue temperature in a range from -50°C to +15°C, - 40°C to +10°C, -35°C to 5°C , -30°C to 0°C or -20°C to -5°C.

In one embodiment, the device may be configured to cool the at least one cryo needle in the cryo needle cooling area such that a resulting tissue cooling zone is generated, in particular in subcutaneous (white) fat tissue, in which everywhere the temperature is lower than a first temperature, wherein the first temperature may in particular be +15°C, +14°C, +13°C, +12°C, +11 °C, +10°C, +9°C, +8°C, +7°C, +6°C, +5°C, +4°C, +3°C, +2°C,

+1 ° or 0°C, but not lower than -50°C, -40°C, -39°C, -38°C, -37°C, -36°C, -35°C, -34°C, -

33°C, -32°C, -31 °C,-30°C, -29°C, -28°C, -27°C, -26°C, -25°C, -24°C, -23°C, -22°C, - 21°C, -20°C, -19°C, -18°C, -17°C, -16°C, -15°C, -14°C, -13°C, -12°C, -11 °C, -10°C, -9°C, -8°C, -7°C, -6°C, -5°C, -4°C, -3°C, -2°C or -1°C.

In a preferred embodiment, the device may be configured for generating different tissue cooling zones, in particular different in shape, size and/or temperature distribution within the tissue cooling zone, depending on the location and/or amount of fat tissue to be treated by cryolipolysis with said device.

In an advantageous embodiment, the device may in particular be configured to establish a tissue cooling zone in subcutaneous fat tissue, within the fat cells are cold down to a tissue temperature in a range from -50°C to +15°C, -40°C to +10°C, -35°C to 5°C, -30°C to 0°C or -20°C to -5°C.

To form a suitable tissue cooling zone, in particular an advantageous tissue cooling zone, preferably a tissue cooling zone as described above, in at least one embodiment, a device according to the present invention may be configured to apply defined amount of cold to a target tissue cooling zone.

In one possible embodiment, a device according to the present invention may in particular be configured for the application of a defined cold in the range from +4°C to -30°C for a defined time to a tissue volume surrounding the cryo needle tip of said device, wherein the device may preferably be configured for application of cold in a range from 0°C to -25°C or form 0°C to -20°C. The device may in particular be configured for application of a defined cold for a time in a range from 10 sec to 10 min, 15 min or 20 min.

In one possible embodiment, a device according to the present invention may in particular be configured for the application of a defined cold in a range between a first temperature and a second temperature, wherein the first temperature may in particular be 4°C, 3°C, 2°C, 1 °C, 0°C, -1°C, -2°C, -3°C, -4°C, -5°C, -6°C, -7°C, -8°C, -9°C, -10°C, -11 °C, -12°C, - 13°C, -14°C or -15°C. The second temperature may for example be -1 °C, -2°C, -3°C, - 4°C, -5°C, -6°C, -7°C, -8°C, -9°C, -10°C ,-11 °C, -12°C, -13°C, -14°C, -15°C, -16°C, -17°C, -18°C, -19°C, -20°C, -21 °C, -22°C, -23°C, -24°C or -25°C, wherein the second temperature may in particular be at least 1 K, 2K, 3K, 4K or 5K respectively 1 °C, 2°C, 3°C, 4°C or 5°C lower than the first temperature.

In one embodiment of a device according to the present invention, the device may be configured for application of a defined cold to a tissue surrounding the cryo needle within a temperature range as described above, preferably in a range from +4°C or 0°C to -20°C, -25°or -30°C, wherein the device may more preferably be configured such that the defined cold can be adjusted within this range in 1 °C or 2°C steps.

In one possible embodiment, a device according to the present invention may in particular be configured for the application of cold in the range between a first application time and a second application time, wherein the first application time may in particular be 2s, 3s, 4s, 5s, 6s, 7s, 8s, 9s, 10s, 11s, 12s, 13s, 14s, 15s, 16s, 17s, 18s, 19s, 20s, 21s, 22s, 23s,

24s, 25s, 26s, 27s, 28s, 29s, 30s, 31s, 32s, 33s, 34s, 35s, 36s, 37s, 38s, 39s, 40s, 41s,

42s, 43s, 44s, 45s, 46s, 47s, 48s, 49s, 50s, 51s, 52s, 53s, 54s, 55s, 56s, 57s, 58s, 59s,

60s (=1 min), 75s, 90s, 105s, 120s (=2 min), 135s, 150s, 165s, 180s (=3 min), 195s, 210s,

225s, 240s (=4 min), 255s, 270s, 285s, 300s (=5 min), 330s, 360s (=6 min), 390s, 420s (=7 min), 450s, 480s (=8 min), 510s, 540s (=9 min), 570s or 600s (=10 min) or 11 min, 12 min, 13 min, 14 min or 15 min.

The second application time may for example be 7s, 8s, 9s, 10s, 11s, 12s, 13s, 14s, 15s, 16s, 17s, 18s, 19s, 20s, 21s, 22s, 23s, 24s, 25s, 26s, 27s, 28s, 29s, 30s, 31s, 32s, 33s, 34s, 35s, 36s, 37s, 38s, 39s, 40s, 41s, 42s, 43s, 44s, 45s, 46s, 47s, 48s, 49s, 50s, 51s, 52s, 53s, 54s, 55s, 56s, 57s, 58s, 59s, 60s (=1 min), 75s, 90s, 105s, 120s (=2 min), 135s, 150s, 165s, 180s (=3 min), 195s, 210s, 225s, 240s (=4 min), 255s, 270s, 285s, 300s (=5 min), 330s, 360s (=6 min), 390s, 420s (=7 min), 450s, 480s (=8 min), 510s, 540s (=9 min), 570s or 600s (=10 min) or 11 min, 12 min, 13 min, 14 min, 15 min, 16 min, 17 min, 18 min, 19 min or 20 min, wherein the second temperature may in particular be at least 5s more than the first application time.

In one embodiment of a device according to the present invention, the device may be configured for application of a defined cold to a tissue surrounding the cryo needle tip within a time range as described above, preferably in a range from 2s, 5s, or 10s to 10 min, 15 min or 20 min, wherein the device may more preferably be configured such that the defined application time can be adjusted within this range in 1s, 2s, 5s, 10s, 20s or 30s steps.

In a preferred embodiment of a device according to the present invention, the device may be configured for application of a defined cold to a tissue surrounding the cryo needle within a temperature range as described above, preferably in a range from +4°C or 0°C to -20°C, -25°or -30°C, wherein the device may more preferably be configured such that the defined cold can be adjusted within this range in 1 °C or 2°C steps, and also for application of the defined cold within a time range as described above, preferably in a range from 2s, 5s or 10s to 10 min, 15 min or 20 min, wherein the device may more preferably be configured such that the application time can be adjusted within this range in 1s, 2s, 5s, 10s, 20s or 30s steps.

In an advantageous embodiment, the device may be configured to establish a tissue cooling zone in subcutaneous fat tissue of a defined size, in particular at least of a first size. For example, the device may be configured to establish a subcutaneous cooling zone of a first size having a volume in a range of 0.5 cm³ up to 10cm³. In particular the device may be configured to generate a tissue cooling zone in subcutaneous fat tissue of a first size having volume of at least 0.5 cm^3, 1 cm³, 1.5 cm³, 2 cm³, 2.5 cm³, 3 cm³, 3.5 cm³, 4 cm³, 4.5 cm³, 5 cm³, 6 cm³, 7 cm³, 8 cm³, 9 cm³ or 10cm³.

In one embodiment, the device may be configured to generate subcutaneous cooling zones of different sizes and/or shapes, in particular depending on the respective use case, in particular for cryolipolysis of subcutaneous fat tissue at different locations in a human body (face, belly/abdomen, legs, arms, back, buttock, calf) and/or penetration depth. The device may for example be configured to generate a tissue cooling zone of a first size and shape in a first cooling cycle and of a second size and/or shape in a second cooling cycle.

In one embodiment, the shape, size and/or temperature distribution within the tissue cooling zone can be adapted, in particular adjusted to the respective use case for advantageous cryolipolysis results. In particular, the defined temperature and the course of the isothermal of said defined temperature, which defines the boarders of the cooling zone to the surrounding subcutaneous tissue, can be adjusted or set, in particular with respect to the location of the subcutaneous tissue to be treated by cryolipolysis using the device.

In one possible embodiment, at least one cryo needle may comprise a thermal isolation and/or comprise a thermal isolation material or may be made at least partly of a thermal isolating material. By such a thermal isolation material, the design and location of the cooling area(s) and non-cooling area(s) along the needle and/or the shape of the needle cooling zone and/or the amount of cooling introduced into the surrounding tissue and the tissue cooling zone can be influenced, in particular adapted. By variation of the thermal isolation properties of the isolation material and/or and its thickness, its arrangement and/or its shape, the location, the shape and the size of the resulting needle cooling zone and/or the resulting temperature distribution within the needle cooling zone and therewith also the location, the shape and the size of the resulting tissue cooling zone and/or the resulting temperature distribution within the tissue cooling zone can be adapted.

In one embodiment, a layer of thermal isolation material may for example be arranged under the outer surface of the cooling needle, in particular in a proximal section of the needle, to reduce the cooling of the outer surface of the needle in this area and to avoid cold burn in the penetration area of the skin or other tissue contacting the outer needle surface, during the cryolipolysis process when the needle penetrates the skin.

In one embodiment, no thermal isolation material may be arranged in the area around the distal needle end, to allow a maximum cooling of subcutaneous tissue, in particular of subcutaneous fat cells, surrounding the needle tip area during the cryolipolysis process. However, in case the generation of a needle and/or tissue cooling zone with lower temperatures is wanted, the presence of a thermal isolating material at the distal needle end, can be advantageous. In addition or alternatively, a needle material with a different thermal insulation may be chosen and/or used.

For defined application of a defined amount of cold, a device according to the present invention may in particular be configured to generate a defined needle cooling temperature, i.e., a defined temperature of or a defined temperature at the outer surface of the cooling needle, in particular of or at the outer surface of the cryo needle in at least one zone of the cryo needle cooling area, in particular in the complete needle cooling area.

The device may in particular be configured to cool down the outer surface of the cryo needle at least at some point or in some portion of the needle cooling area to +14°C, +13°C, +12°C, +11 °C, +10°C, +9°C, +8°C, +7°C, +6°C, +5°C, +4°C, +3°C, +2°C, +1 ° or

0°C, but not lower than -51 °C, -50°C, -40°C, -39°C, -38°C, -37°C, -36°C, -35°C, -34°C, -

33°C, -32°C, -31°C,-30°C, -29°C, -28°C, -27°C, -26°C, -25°C, -24°C, -23°C, -22°C, -21 °C,

-20°C, -19°C, -18°C, -17°C, -16°C, -15°C, -14°C, -13°C, -12°C, -11 °C, -10°C, -9°C, -8°C, -7°C, -6°C, -5°C, -4°C, -3°C, -2°C or -1°C.

In one embodiment of a device according to the present invention, the device may be configured to cool down the outer surface of the cryo needle at least at some point or in some portion of the needle cooling area to an outer surface temperature in a range from +4°C or 0°C to -20°C, -25°or -30°C, wherein the device may more preferably be configured such that said outer surface temperature can be adjusted within this range in 1 °C or 2°C steps.

In one embodiment of a device according to the present invention, the device may be configured for application of a defined cold to a tissue surrounding the cryo needle tip within a time range as described above, preferably in a range from 2s, 5s or 10s to 10 min, 15 min or 20 min, wherein the device may more preferably be configured such that the application time (of the cold) can be adjusted within this range in 1s, 2s, 5s, 10s, 20s or 30s steps.

In a preferred embodiment of a device according to the present invention, the device may be configured for application of a defined cold to a tissue surrounding the cryo needle within a temperature range as described above, preferably in a range from +4°C or 0°C to -20°C, -25°or -30°C, wherein the device may more preferably be configured such that the defined cold can be adjusted within this range in 1 °C or 2°C steps, and also for application of the defined cold within a time range as described above, preferably in a range from 2s, 5s or 10s to 10 min, 15 min or 20 min, wherein the device may more preferably be configured such that the application time (of the cold) can be adjusted within this range in 1s, 2s, 5s, 10s, 20s or 30s steps. In a preferred embodiment, the cooling of the outer needle surface in the needle cooling area may be limited, for example to a defined temperature limit value and/or a defined application time limit, wherein the temperature limit value and/or the application time limit may in particular be adjusted and/or set by a practitioner, for example by appropriate adjustment of one or more control parameters and/or by selection of a specific cryo needle assembly. The limit may in at least one embodiment of a device according to the present invention depend on one or more parameters. For example, the limit may depend on the applied cold or cooling temperature so far and/or on the passed application time and/or a combination thereof. This allows precise control of the cooling of the subcutaneous tissue. Precise control and adjustment of the cooling of the subcutaneous tissue seems in particular to be important for fat cells to achieve a controlled cell death of adipocytes with as few side effects as possible.

In one possible embodiment, the device may comprise one or more valves to control, in particular to limit and/or to control, a cooling flow of the cryo fluid into and/or through the at least one cryo needle. Such a valve may contribute significantly to the control of the cooling process. When using a precise and fast reacting valve which opens and closes precisely and immediately in response to a user’s command, precise cooling of subcutaneous tissue, in particular of subcutaneous fat tissue, can be achieved.

In one embodiment, at least one valve may be configured to mix a cooling flow with ambient air for improved control and/or adjustment of the cooling flow and/or for advantageous adjustment of the cooling temperature, in particular for improved control of the cooling temperature, which occurs on the outer surface of the needle when cryo fluid is flowing through the cryo needle.

In one possible embodiment, the device may be a handheld device, similar to a dental device. In a preferred embodimentthe weight of the handheld device is below 0.5kg, 0.4kg, 0.3kg or 0.2kg. The lighter the device is, the less tiring is its use for the practitioner. For precise treatment it seems to be important that the device is well-balanced and can be hold in the hand without requiring a counter momentum to be applied by the holder. In particular, for comfortable and precise use, the device should neither be top-heavy, i.e. it should not tend to tilt away from the needle tip when in use, nor "tail-heavy", i.e. it should not tend to tilt downwards with its end facing away from the needle tip. To provide a well- balanced handheld device, which is neither top-heavy nor tail-heavy, the device may comprise one or more counter masses or regions of counter mass concentration. A counter mass might, for example, and in particular be helpful in case the device comprises a cryo fluid cartridge which is arranged at the proximal end of the device (at the “tail-end”).

To achieve a comfortable and well-balanced handheld device, the parts with the highest weight may preferably be arranged in a centre area of the main body, in particular in or near a gripping area. In one embodiment of a handheld device configured for holding a cryo fluid cartridge, in particular the cartridge may be arranged in a centre area of the main body and/or near the centre of gravity of said handheld device.

In one possible embodiment, the main body may be couplable to a stationary cryo fluid storage as cryo fluid source, preferably of CO2 or N2 or N2O or a combination thereof, in particular via at least one supply line. The main body may in particular be coupled to a cryo fluid source by one or more supply lines, hoses, or tubes. The device may comprise at least a first hose or tube or supply line for a flow of cryo fluid, in particular cryo gas, from a cryo fluid source to the main body, preferably through the main body, and further into the cryo needle. In one embodiment, the device may also comprise a second hose or tube or return line for a flow of cryo fluid, in particular cryo gas, from the cryo needle backwards. The return flow may be directed into the environment or may be recycled at least partially and directed back into a cryo fluid flow circulation. The return flow may also be directed through the main body.

The stiffness of such a supply line or return line and/or hose and/or tube may in particular be as low as necessary, to affect the handling as less as possible. Ideally, the supply line is almost not be noticed during use and handling feels like using a tubeless mobile device.

In addition or alternatively, the device, in particular the main body, may be couplable to a cryo fluid cartridge as cryo fluid source, in particular to a cryo fluid cartridge being filled with CO2 or N2 or N2O or a combination thereof. How such a coupling can be realized, is for example known from the iovera® device. The cryo fluid may in particular be a medical fluid which is appropriate for medical use and comprises a sufficient pureness, in particular sufficient medical purity.

In one possible embodiment, the cryo needle assembly may be interchangeably and detachably coupled or couplable to the main body, wherein in particular the main body and/or the cryo needle assembly may comprise an adapter, via which the cryo needle assembly and the main body are coupled or can be coupled to each other. An adapter allows an easy and flexible coupling of different cryo needle assemblies to the main body. Thereby flexibility of the device can be increased quite easy. Further, an adapter may allow in a quite easy manner to realize different orientations of a longitudinal axis of the cryo needle relative to the main body. This allows adaptation of the handling properties of the device and to achieve an improved and/or more precise handling of the device in at least some cases. In particular, in some cases and by using an appropriate adapter, accessibility of certain parts of the body can be improved.

In one possible embodiment, the at least one cryo needle may be a 15G, 16G, 17G, 18G, 19G, 20G, 21 G, 22G, 23G, 24G, 25G, 26G, 27G, 28G, 29G, 30G, 31 G, 32G, 33G, 34G or 35G needle or cannula. It is only important that a sufficient cooling can be achieved, in particular a sufficient cooling based on the Joule-Thomson-Effect as it known from cryosurgery.

The device may in one embodiment in particular be configured for use with different cryo needles and/or different cryo needle assemblies, wherein the device may in particular be configured for use with different cryo needle assemblies, which are adapted for different treatment areas, as for example for percutaneous cryolipolysis of subcutaneous fat cells (adipocytes and/or preadipocytes) in the face (chin, cheeks), at the thighs, the calf, the hips, the abdomen, the butt, the arms, the breast, the upper back and/or the upper back. This allows, for example, in regions with higher amounts of subcutaneous fat tissue, the use of larger cryo needle sizes, which results in increase of efficiency of the cryolipolysis treatment since a higher volume of fat tissue can be destroyed in the same or less time. In regions with less amount of subcutaneous fat tissue volume or in regions where very precise punctuation and placement of the cryo needle is important to avoid a damage of surrounding structures as vessels, nerves, organs, muscles and/or lymphatics, a smaller cryo needle can be used for more precise and therewith safer cryolipolysis.

In one possible embodiment, the needle assembly may comprise at least two cryo needles, in particular a group of needles, in particular an array of cryo needles. Alternatively or in addition, the needle assembly may comprise one or more micro needles, in particular at least one micro needle array.

A “needle array” in the meaning of the present disclosure means a group of needles, wherein the needles are arranged relatively to each other geometrically defined, in particularly according to a defined or regular pattern.

A “group of needles” means a geometrically undefined arrangement of one or more cryo needles to each other.

A “micro needle” means a needle which is shorter and/or thinner as a usual needle with the same size.

The use of a group or array of cryo needles may in particular be advantageous in regions with higher amounts of subcutaneous fat tissue, wherein the use of micro needles and/or of a cryo needle assembly comprising only a single cryo needle seems to be advantageous in regions with less amount of subcutaneous fat tissue or in regions where very precise punctuation and placement of the cryo needle is important to avoid a damage of surrounding structures as vessels, nerves, organs, muscles and/or lymphatics.

In one embodiment, the device may be configured to generate subcutaneous cooling zones of different sizes and/or shapes for the different needles of the needle group, in particular depending on the respective use case, in particular for cryolipolysis of subcutaneous fat tissue at different locations in a human body (face, belly/abdomen, legs, arms, back, buttock, calf) and/or penetration depth. The device may for example be configured to generate a tissue cooling zone of a first size and shape adjacent to a first needle and of a second size and/or shape adjacent to a second needle. To keep sufficient distance to organs and other structures like muscles and main vessels and nerves, one or more markers (similar to a scale) may be provided on the outer needle surface to indicate penetration/injection depth.

In one embodiment alternatively and/or in addition, the cryo needle assembly, in particular the cryo needle, may comprise an end stop, which may contact the skin surface as soon as the permissible penetration depth is reached and blocks deeper penetration.

In one possible embodiment, the device may comprise a control unit configured for controlling and/or adjustment and/or setting of the cooling of the cryo needle cooling area, in particular for controlling, adjusting and/or setting of a cooling temperature in the cryo needle cooling area, to control the cooling temperature in the tissue cooling zone. In a preferred embodiment, the device may be configured such a desired tissue cooling zone may be generated, wherein in particular one or more parameters may be set and/or adjusted before or during the cryolipolysis process to adapt the tissue cooling zone properties to the specific treatment.

In one embodiment, one or parameters may be adjusted and/or set, in particular in dependence of the location of subcutaneous fat tissue to be treated by cryolipolysis, the volume of fat tissue to be treated, the average size of the fat cells to be treated and the indication for treatment (lipoedema, lipoma, cosmetic reasons, aesthetic reasons).

In one embodiment the device may in particular be configured such that a cooling temperature of the needle cooling zone, in particular the temperature at a defined point on the outer needle surface, preferably the cooling temperature distribution over the outer needle surface in the needle cooling area, may be adjusted, wherein cooling can preferably be caused as adjusted. This allows an optimal adaptation of the needle cooling area to several different use case requirements.

The device may comprise at least one temperature sensor, in particular a temperature sensor to detect a tissue temperature and/or a temperature sensor for detecting the cryo needle(s)’s surface temperature. In one embodiment, the device may comprise a display unit to present the cryo needle(s)’s surface temperature and/or a passed and/or a remaining application time and/or at least one other relevant parameter to the practitioner. The device may issue an alert, when the temperature exceeds an appropriate range or exceeds a predetermined threshold and/or when a defined application time is reached, for example. The device may indicate start and/or end of the cooling and/or indicate that the needle may be removed from the subcutaneous fat tissue.

In one embodiment the device may in particular be configured such that a cooling temperature of the tissue cooling zone, in particular the defined temperature, preferably the cooling temperature distribution in the tissue cooling zone, and/or a needle cooling temperature (cooling temperature at an outer surface of the cryo needle), and/or a defined application time may be adjusted, wherein cooling can preferably be caused as adjusted. This allows an optimal adaptation of the tissue cooling zone to several different use case requirements.

In one embodiment, the device may in particular be configured such that a target volume of fat tissue can be set, for which apoptosis and/or necrosis should be initiated, and/or a defined target volume ratio of cell death caused by apoptosis to cell death caused by necrosis.

In one embodiment, in particular a defined application time can be set and/or a defined tissue cooling temperature and/or a defined tissue cooling volume and/or a defined needle cooling temperature (outer needle surface temperature).

The device may comprise one or more displays, in particular to indicate the status of the device and/or to indicate the status of the cooling process or, for example of the tissue temperature, and/or a passed and/or a remaining application time and/or at least one other relevant parameter to the practitioner. In one embodiment, the device may be configured to visualise the adjusted/set tissue cooling zone and/or the generated tissue cooling zone.

The device may be programmable. The device may be configured to store predetermined treatment programs for cryolipolysis treatment for different use cases comprising appropriate default parameters and/or treatment schemes or patterns. The device may be configured to guide the practitioner through a treatment process.

The device may comprise a timer, in particular for precise control of the cooling time. In a preferred embodiment, the device may be configured such that the cooling temperature may be adjusted and/or controlled over time. In particular one or more time dependent cooling temperature profiles can be set.

In one embodiment, the device may be configured for cooling of subcutaneous fat tissue with a cooling time, in particular in combination of any of the cooling temperatures and/or temperature ranges as described herein, in a range from 1s, 2s, 3s, 4s, 5s, 6s, 7s, 8s, 9s, 10s, 11s, 12s, 13s, 14s, 15s, 16s, 17s, 18s, 19s, 20s, 21s, 22s, 23s, 24s, 25s, 26s, 27s, 28s, 29s, 30s, 31s, 32s, 33s, 34s, 35s, 36s, 37s, 38s, 39s, 40s, 41s, 42s, 43s, 44s, 45s, 46s, 47s, 48s, 49s, 50s, 51s, 52s, 53s, 54s, 55s, 56s, 57s, 58s, 59s, 60s (=1 min), 75s, 90s, 105s, 120s (=2 min), 135s, 150s, 165s, 180s (=3 min), 195s, 210s, 225s, 240s (=4 min), 255s, 270s, 285s, 300s (=5 min), 330s, 360s (=6 min), 390s, 420s (=7 min), 450s, 480s (=8 min), 510s, 540s (=9 min), 570s or 600s (=10 min) to 20 min, 19 min, 18 min, 17 min, 16 min, 900s (=15 min), 870s, 840s (=14 min), 810s, 780s(=13 min), 750s, 720s (=12 min), 690s, 660s (=11 min), 600s (=10 min), 570s, 540s (=9 min), 510s, 480s (=8 min), 450s, 420s (=7 min), 390s, 360s (=6 min), 330s, 300s (=5 min), 285s, 270s, 255s, 240s (=4 min), 225s, 210s, 195s, 180s (=3 min), 165s, 150s, 135s, 120s (=2 min),

105s, 90s, 75s, 60s (=1 min), 59s, 58s, 57s, 56s, 55s, 54s, 53s, 52s, 51s, 50s, 49s, 48s,

47s, 46s, 45s, 44s, 43s, 42s, 41s, 40s, 39s, 38s, 37s, 36s, 35s, 34s, 33s, 32s, 31s, 30s,

29s, 28s, 27s, 26s, 25s, 24s, 23s, 22s, 21s, 20s, 19s, 18s, 17s, 16s, 15s, 14s, 13s, 12s,

11s, 10s, 9s, 8s, 7s, 6s or 5s.

A kit for percutaneous cryolipolysis according to the present invention comprises a device and a cryo fluid source, wherein the device comprises a main body, to which a cryo fluid source is couplable, and a cryo needle assembly, being coupled or couplable to the main body. The cryo needle assembly comprises at least one cryo needle, wherein the least one cryo needle is configured to penetrate the skin of a human or an animal at least partially. The device is configured to cause a cooling of at least one cryo needle at least in a cryo needle cooling area, when a cryo fluid source and the cryo needle assembly are coupled to the main body, by using cryo fluid from the cryo fluid source, and to form a subcutaneous tissue cooling zone adjacent to the cryo needle cooling area of the cooled cryo needle, when the cooled cryo needle is penetrating the skin of a human or an animal at least partially. The device is further configured to cool subcutaneous fat cells located in the tissue cooling zone such that at least for some of the fat cells located in the tissue cooling zone apoptosis and/or necrosis is initiated.

The device of the kit may in particular be a device according to the present invention as described above. The device and the cryo fluid source may in particular be coupled, in particular such that they are in fluid communication, in particular such that cryo fluid from the cryo fluid source can be flow into and/or through the cryo needle.

According to the present invention, a device configured for percutaneous cryolipolysis or a kit configured for percutaneous cryolipolysis as described above may in particular be used for cosmetic, aesthetic or therapeutic application, particularly for percutaneous cryolipolysis.

A device or a kit according to the present invention may in particular be used for subcutaneous cryolipolysis in combination with one or more defined treatment schemes or patterns and/or according to different instructions for use and/or for different indications. Such schemes may define allowed and not allowed zones for treatment and for example penetration directions, penetration depth, cooling temperature and time, to reduce the risk of complications and unwanted side effects.

A device or kit according to the present invention may in particular be used for percutaneous cryolipolysis of lipoedema, lipoma and/or for cosmetic aesthetic or therapeutic applications of human or animal fat tissue as for example for percutaneous cryolipolysis of subcutaneous fat cells (adipocytes and/or preadipocytes) in the face (chin, cheeks), at the thighs, the calf, the hips, the abdomen, the butt, the arms, the breast, the upper back and/or the upper back.

In one embodiment the device and or the kit may be configured such that the percutaneous cryolipolysis may be performed under image control (e. g. ultrasonic, MRT, CT). In some cases it might be advantageous, to apply a local anesthetic composition to the area where the skin is to be pierced first and before the skin is pierced, to make the treatment more comfortable for the patient. For example, a cream containing lidocaine or another local anesthetic composition may be applied first. In some cases, for piercing of the skin, in particular if the cryo needle is a blunt needle or cannula, another means may be used. That means in other words, after the optional local anesthetic has been applied, the skin may be pierced by a piercing means first, before the cryo needle is injected through the pierced skin and placed with its needle cooling area into the subcutaneous fat tissue.

A device or a kit according to the present invention may also be used instead of liposuction. It seems that by using percutaneous cryolipolysis a safer reduction of body fat tissue with less risks for the patient may be possible. However, it has to be noted that the amount of subcutaneous fat tissue for which death by apoptosis and/or necrosis is caused by percutaneous cryolipolysis might be limited by the abilities of the body to break down the dead fat cells.

The present invention described above will now be further explained by the following, nonlimiting embodiments and examples, wherein further optional features of the inventions are disclosed in the drawings and in the description of these drawings. All features and all combinations of features described above and/or outlined below and/or only illustrated in the drawings can be realized in an embodiment of the invention in the combination described or stand alone or in at least one other combination not described explicitly herein, as this combination is technically reasonable.

BRIEF DESCRIPTION OF THE DRAWINGS

The accompanying drawings, which are incorporated herein and form a part of the specification, illustrate schematically preferred embodiments of the inventions described above, in particular of the present invention, and together with the description, serve to explain the principles and at least partly the features of the described inventions, wherein for same parts same reference signs are used. It is illustrated in: Fig. 1 a perspective view of an example of a kit for percutaneous cryolipolysis according to the present invention with a first example of a device for percutaneous cryolipolysis according to the present invention,

Fig. 2 a perspective view of a second example of a device for percutaneous cryolipolysis according to the present invention,

Fig. 3 a schematic illustration of a portion of the cryo needle assembly of the device of the kit as illustrated in Fig. 1 during use while penetrating the skin of a human and cooling of subcutaneous fat cells to cause subcutaneous cryolipolysis and fat cell death by apoptosis and/or necrosis,

Fig. 4 an enlarged schematic illustration of a first exemplary interior design of the cryo needle of Figs. 1 to 3 in its tip region in the cooling area,

Fig. 5 an enlarged schematic illustration of a second exemplary interior design of the cryo needle of Figs. 1 to 4 in its tip region in the cooling area,

Fig. 6 exemplary body regions for possible use of a kit or device according to the present invention for percutaneous cryolipolysis,

Fig. 7 a side view of a third example of a device for percutaneous cryolipolysis according to the present invention,

Fig. 8 a perspective view of a fourth example of a device for percutaneous cryolipolysis according to the present invention,

Fig. 9 another enlarged schematic illustration of a further exemplary design of the cryo needle of Figs. 1 to 4 in its tip region in the cooling area,

Fig. 10 bright field microscopic images of results from experiments with adipocytes, to which a defined cold has been applied for a defined time under defined conditions,

Fig. 11 fluorescence image analysis of cell nuclei of adipocytes stained in blue to which a defined cold has been applied for a defined time under defined conditions, Fig. 12 a diagram showing the resulting nuclei counts of the fluorescence image analysis of the adipocytes illustrated in Fig. 11 depending on the temperature of the cooling temperature the cells had been subjected to, and

Figs. 13, 14 results from Lactate Dehydrogenase (LDH) assay tests to assess cytotoxicity of temperature treatments applied to adipocytes under different conditions.

Fig. 1 shows a perspective view of an example of a kit 100 for percutaneous cryolipolysis according to the present invention with a first example of a device 10 for percutaneous cryolipolysis according to the present invention and a cryo fluid source 15.

The device 10 is an elongated handheld device 10 similar to a common dental device and comprises a torpedo-shaped main body 13 and a cryo needle assembly 11 comprising a needle hub 20 and a cryo needle 12.

The main body 13 comprises a proximal end and a distal end (not indicated with reference signs), wherein at the proximal end of the main body 13 the cryo needle assembly 11 is detachably mounted to the main body 13. The main body 13 is configured to support several different cryo needle assemblies 11. At the distal end of the main body 13 a first end of a supply line 14 for supplying the cryo needle assembly 11 with a cryo fluid is connected. The second end of the supply line 14 is connected, in particular fluidly coupled to the cryo fluid source 15 in form of a stationary cryo gas container 15 having a volume of 25I filled with carbon dioxide (CO2) as cryo gas.

The device 10 further comprises a control unit 16 integrated into a housing, in which also the cryo fluid source 15, in particular the gas container filled with carbon dioxide (CO2), is arranged. The housing and the control unit 16 are part of an operation terminal by which the kit 100, in particular the device 13, can be operated and controlled by a practitioner. Similar to a dental device. The operation terminal comprises several operational control buttons 17 and may further comprise one or more displays (not shown) to indicate the status of the kit 100 and/or the device 13. Instead of control of the operation terminal by buttons 17, controlled by foot switch (not shown) may be possible. In the illustrated example, the cryo needle assembly 11 comprises a straight needle hub 20 and a 25G straight cryo needle 12, wherein the needle hub 20 and the cryo needle 12 are arranged with their longitudinal axis along a common axis, which also coincides with a longitudinal axis of the main body 13.

The main body 13 is configured such that cryo fluid from the cryo gas source 15 may be directed via the supply line 14 and through the main body 13 into the cryo needle assembly 11 , in particular almost to the needle tip of cryo needle 12, to cause a cooling of the outer surface of the cryo needle 12 in a needle cooling area P1 (see Fig. 3-5) based on the Joule-Thomson-Effect, and as it is in general known from cryosurgery devices for cryotherapy of tumorous tissue.

The device 10 may further comprise a gripping area at the main body for comfortable and ergonomic handling and a secure grip by a practitioner. For precise treatment of subcutaneous fat tissue by percutaneous cryolipolysis, the device 10 is further in particular well-balanced to avoid tilting in a practitioner’s hand.

In other embodiments the needle hub 20 and/or the cryo needle 12 may be angled or may be arranged with an angle relative to the longitudinal axis of the main body 13, as it is, for example, illustrated in Fig. 2, which shows a perspective view of a second example of a device 10A for percutaneous cryolipolysis according to the present invention. A design as illustrated in Fig. 2 might in some use cases be more advantageous, since a different accessibility compared to the straight design as illustrated in Fig. 1 may be achieved.

Another difference between device 10 in Fig. 1 and device 10A in Fig. 2 is that the device 10A in Fig. 2 in addition and compared to the device 10 in Fig. 1 comprises an adapter 19 between the main body 13 and the needle hub 20. The angle between the longitudinal axis of the cryo needle 12 and the longitudinal axis of the main body 13 is in particular caused by said adapter 19. By changing the adapter 19, several configurations for different use cases can easily be realized. This makes the device 10A quite flexible, in particular more flexible than the device in Fig 1 , for percutaneous cryolipolysis in several body regions. The device in figure 2 further comprises a supply line connection portion 18, which has a similar function as the adapter 19 between the main body and the needle hub 20, just with respect to the supply line 14.

According to the present invention, both devices 10 and 10A as illustrated in Fig. 1 and 2 are configured for percutaneous cryolipolysis of subcutaneous fat cells (adipocytes and/or preadipocytes).

Fig. 3 shows a schematic illustration of a portion of the cryo needle assembly 11 of the device 10 of kit 100 as illustrated in Fig. 1 during use. As illustrated in Fig. 3, the cryo needle 12 is configured to penetrate the skin with its dermis layers D1 and D2 and to be placed at least partly, in particular with its tip area and its needle cooling area P1 , in the subcutaneous fat tissue layer F to cool fat cells A (adipocytes and/or preadipocytes) surrounding the needle cooling area P1.

By using cryo fluid G from the cryo fluid source 15 and directing a cryo fluid flow into the cryo needle 12 (flow G1 ) and back (return flow G2), an outer surface of the needle 12 can be cooled in the needle cooling area or portion P1. This results in the generation of a cooling zone CZ in the surrounding subcutaneous fat tissue F, which has an oval or egg shape in the illustrated example.

To protect in particular the dermis layers D1 and D2 of the skin above the subcutaneous fat tissue F, the portion P2 of the cryo needle 12 is a non-cooling area P2, in which no cooling of the outer needle surface is caused to avoid cold burn of the tissue adjacent and/or surrounding needle portion P2. Cooling of the outer needle surface is only caused in portion P1 (the needle cooling area) in the needle tip area.

By the cryo needle 12 subcutaneous fat cells A can be cold such that cell death in initiated. The device 13 is in particular configured such that at least for some of the fat cells A located in the tissue cooling zone CZ apoptosis and for others necrosis is initiated, wherein cell death by apoptosis is indicated by the symbol with the white cross with reference sign A1 and cell death by necrosis is indicated by the symbol with the black cross with reference sign A2. Other structures as for example muscles M or deeper layers are preferably not affected by the cooling.

The tissue cooling zone CZ, in particular its size, shape and/or dimensions, is defined by the isothermal of a first temperature T 1 , which may for example be T1 = -1 °C. Everywhere within the tissue cooling zone, the tissue temperature is -1 °C or lower. For visualisation of the temperature distribution, the isothermals of a second temperature (e.g. T2 = -5°C) and a third temperature (e.g. T3 = -10°C) are also illustrated schematically.

With a cooling zone CZ according to Fig. 3, a cell death ratio of 1 :1 can be achieved, wherein 100% of the fat cells in the cooling zone CZ died. 50 Vol.% of the fat cells died caused by apoptosis A1 and 50 Vol% of the fat cells by necrosis A2.

The flow of the cryo fluid G in the interior of the needle 12 is illustrated in more detail in Fig. 4, which shows an enlarged schematic illustration of a first exemplary interior design of the cryo needle of Figs. 1 to 3 in its tip region 21 in the cooling area.

In this example of a cryo needle 12 the cryo fluid flow G1 from the cryo fluid source 15 is directed into inner lumen 12A and flow through said lumen 12A along the center axis of the cryo needle 12 almost to the closed needle tip 21. The tip 21 is closed to avoid leakage or transmission into surrounding tissue. At the end of the inner lumen 12A the cryo fluid flow is returned and flows back as flow G2 through lumen 12B, thereby cooling the outer surface of the cryo needle 12 and the cryo fluid in the inner lumen 12A to increase the overall cooling.

In other embodiments of a device according to the present invention, flow direction may be vice versa. In another embodiment, one half of the complete inner lumen of the cryo needle might be used for the supply flow G1 and the other half for the return flow G2. Further embodiments are possible.

With the cooling zone CZ according to Fig. 4, which is based on a different device adjustment compared to Fig. 3, a cell death ratio of 70:30 can be achieved, wherein 100% of the fat cells within the cooling zone CZ died. 70 Vol.% of the fat cells died caused by apoptosis A1 and 30 Vol% caused by necrosis A2, as adjusted before starting the cryolipolysis process.

In the present case the cryo needle 12 has a closed needle tip 21 but is still configured for piercing the skin for percutaneous cryolipolysis. Therefore, it is not necessary to perform a separate piercing step by a separate piercing device to pierce the skin first to enable penetration of the skin by the cryo needle 12 before injecting the cryo needle 12.

Fig. 5 shows an enlarged schematic illustration of a second exemplary interior design of the cryo needle 12 of Figs. 1 to 4 in its tip region 21 in the cooling area P1 , wherein this example of a cryo needle 12 comprises an inner insulating layer 22 to avoid cooling of the outer needle surface in the non-cooling area P2. Thereby, the risk of cold burn of the upper skin layers, for example of dermis layers D1 und D2 (see Fig. 3) can significantly be reduced.

With the cooling zone CZ according to Fig. 5, which is based on the same adjustment as in Fig. 3, a cell death ratio of 50:50 can be achieved, wherein 100% of the fat cells within the cooling zone CZ died. 50 Vol.% of the fat cells died caused by apoptosis A1 and also 50 Vol% caused by necrosis A2, as adjusted before starting the cryolipolysis process.

Fig. 6 shows exemplary body regions L1 to L13 for possible use of a kit 100 or device 10 according to the present invention for percutaneous cryolipolysis (L1 : face, in particular cheek; L2: chin; L3: neck; L4: Brust; L5: arms; L6: belly/abdomen; L7: hips; L8: thighs; L9: calf; L10: ankle; L11 : upper back, shoulder; L12: lower back; L13: buttock). The kit 100 and/or the device 10, 10A may be adjusted or adapted depending on the region for percutaneous cryolipolysis, wherein in particular an appropriate needle assembly 11 may be chosen and/or appropriate cooling parameters, in particular the cooling temperature, may be adjusted and/or set, preferably over time.

Fig. 7 shows a side view of a third example of a device 10B for percutaneous cryolipolysis according to the present invention, wherein this device 10B has a pistol shape and is no longitudinal device and has no longitudinal main body 13. However, this device 10B also has an adaptor 19. The size of the cryo needle 12 is greater than the size of the devices 10 and 10 of Fig. 1 and 2. The cryo flow can be controlled by lever 23. With this device, in particular high volumes of subcutaneous fat tissue A can be treated, for example in body region L6 (belly/abdomen), body region L8 (thighs) and/or body region L13 (buttock). However, this device 10B, 20 is less precise than the devices 10 and 10A of Fig.1 and 2.

Fig. 8 shows a perspective view of a fourth example of a device 10C for percutaneous cryolipolysis according to the present invention, wherein this device is tubeless and comprises and integrated 0,25l cartridge 24 filled with a cryo gas, here N2O. To achieve a well-balanced handling, into the main body 13 a counterweight 26 is integrated. The device 10C further comprises an end stop 25 at the needle hub, to prevent too deep injection. The device 27 further comprises a status LED and display means for displaying one or more cooling parameters like tissue cooling temperature respectively outer needle temperature and/or cooling time.

Fig. 9 shows another enlarged schematic illustration of a further exemplary design of the cryo needle of Figs. 1 to 4 in its tip region in the cooling area, wherein in this embodiment the device comprises in addition to the device illustrated in Fig. 5 a temperature sensor 29 in the needle tip area. Further, the device illustrated in Fig. 9 is configured such that a needle surface temperature TN of an outer needle surface can be adjusted and be controlled precisely, in particular in 1 °C steps within a range from +4°C to -25°C and for application times/ cooling times in a range from 10s to 15 min in steps of 20s according to the specific needs of the different treatments, for example to achieve a defined and wanted a cell death ratio of approx. 78:12 (cells died by apoptosis : cells died by necrosis) can be achieved.

Fig. 10 shows bright field microscopic images of results from experiments with adipocytes prepared in a defined manner (differentiated from preadipocytes to mature adipocytes within 14 days under defined conditions) after exposure to different conditions, wherein most of the pictures have been taken 12h after end of treatment.

The upper left image shows a first control sample of mature adipocytes prepared as described previously which have been subjected to 37°C for 10 min. instead of being subjected to a defined cooling, wherein the image was captured 12 hours after the end of the treatment. The upper middle image shows a second control sample of mature adipocytes prepared as described previously which have been treated with Staurosporine (a chemical structure by which in a variety of types of cells apoptosis may be initiated or triggered in a defined manner) for 10 min, wherein the image was captured 16 hours after the end of the treatment.

The upper right image shows a sample of mature adipocytes prepared as described previously which have been subjected to a defined cooling of +4°C for 10 min., wherein the image was captured 12 hours after the end of the treatment.

In order to cool the cells in a defined manner, the cells were each provided in a defined state in special cell culture carriers, in this case in so called “Multiwell-Plates”, which are made from plastic and are well-known from prior art. The cell culture carriers with the cells inside were each placed on a cooling plate for the desired, defined cooling time, which was cooled so that it had the wanted, defined cooling temperature on its cooling plate surface.

The bottom left image shows a sample of mature adipocytes prepared as described previously which have been subjected to a defined cooling of +0°C for 10 min., wherein the image was captured 12 hours after the end of the treatment.

The bottom middle image shows a sample of mature adipocytes prepared as described previously which have been subjected to a defined cooling of -20°C for 10 min., wherein the image was captured 12 hours after the end of the treatment.

The bottom right image shows a sample of mature adipocytes prepared as described previously which have been subjected to a defined cooling of -80°C for 10 min., wherein the image was captured 12 hours after the end of the treatment.

Fig. 11 shows fluorescence image analysis of cell nuclei of preadipocytes of different samples stained in blue to which previously the same defined cooling has been applied as in the experiments described in Fig. 10.

The most left image shows the fluorescence image of the first control sample. The image next to the right (indicated with STS) shows the second sample treated with Staurosporine. The next right image shows the stained cell nuclei of the sample treated with +4°C for 10 min. The next right image shows the stained cell nuclei of the sample treated with +0°C for 10 min. The next right image shows the stained cell nuclei of the sample treated with - 20°C for 10 min, and the next right image shows the stained cell nuclei of the sample treated with -80°C for 10 min.

The lower the applied temperature was, i.e., the higher the applied amount of cooling was, the more preadipocytes died, and the less cells have been counted. At -80°C all cells died, and loose cells had been washed out.

Fig. 12 shows a diagram showing the resulting nuclei counts of the fluorescence image analysis of the preadipocytes illustrated in Fig. 11 depending on the temperature of the cooling temperature the cells had been subjected to.

This diagram clearly shows that the lower the applied temperature was, i.e., the higher the applied amount of cooling was, the more preadipocytes died, and the more the cell count decreases. At -80°C all cells died, and loose cells hat been washed out. Cell viability (cell count) may follow a linear decrease with decrease in cooling temperature applied. In a temperature range from +4°C to -20°C, in particular in a temperature range from 0°C to - 20°C, a controlled cell death could be achieved.

Figs. 13 and 14 show results from Lactate Dehydrogenase (LDH) assay tests which have also been done to assess cytotoxicity of temperature treatments applied to adipocytes under different conditions. For this tests mature adipocytes prepared as described above have been used and been analyzed with a LDH assay as generally known.

LDH assay tests are biochemical assays used to quantify the activity of the enzyme lactate dehydrogenase (LDH) in various biological samples, including blood, serum, plasma, and cell cultures. Characteristic for this kind of assay is that LDH is released into the extracellular space when cells are damaged or lysed. The released amount of LDH can be detected and the determined LDH level can serve as a marker for cell death or cell cytotoxity.

Figs. 13 and 14 shows the results of the reaction of different samples of adipocytes which have been subjected to different conditions, wherein Fig. 13 shows the resulting cell reaction after 4 hours of storage of the cells in an incubator subsequently to their defined (cooling) treatment. Fig. 14 shows the resulting cell reaction after 24 hours of storage of the cells in the incubator subsequently to their (cooling) treatment. The samples marked with "ctrl. I", "ctrl. II" and "ctrl. Ill" are each control samples, whereby the first control sample "ctrl. I" is a corresponding positive control sample associated with the assay, which has only been analyzed to check the functionality of the assay kit, but which has not been exposed to a defined temperature and/or cooling.

Control sample II ("ctrl. II") was exposed to a temperature of 37°C for 15 minutes, i.e., to body temperature, and was therefore also not exposed to any cooling.

To prevent the samples from freezing during cooling and the assay from losing its function, it was necessary to preserve the adipocytes that were to be cooled in the tests, for which a 35% glycerol solution was used. The behavior of adipocytes preserved in 35% glycerol exposed to at 37°C for 15 min is shown by the results of control sample III ("ctrl. III").

The samples indicated with 4°C* were preserved in 35% Glycerol and exposed to +4°C for 15 min, the samples indicated with 0°C* were preserved in 35% Glycerol and exposed to 0°C for 15 min, and indicated with -20°C* were preserved in 35% Glycerol and exposed to -20°C for 15 min.

It could be seen that in accordance with the previously described results, the cell cytotoxity, which is inverse to cell viability, decreases with higher cooling amount or decreased cooling temperature respectively. What can also be seen in the experimental data is that after 20h more in the incubator (i.e., after 24h storage time in the incubator compared to 4h storage time in the incubator), cell cytotoxity has further increased without any further additional cooling treatment. This indicates that at least the cells which died additionally during the additional 20h storage in the incubator, died by apoptosis since apoptosis generally occurs with a time delay after the event that triggers cell death, and not by necrosis which generally occurs immediately after the cell death causing event.

Cooling temperatures in a range from +4°C to -25°C, in particular in a range from 0°C to - 20°C, applied to adipocytes and/or preadipocytes seem to be suitable to initiate controlled apoptosis of the respective cells. By variation of the cooling temperature and/or cooling time the cooling energy introduced into the cells/the tissue may be controlled.

In the absence of insulating material between the cells and the cooling surface, as for example in the absence of cell culture carrier plastic material, resulting in direct contact of the cells (adipocytes and/or preadipocytes) with the cooling surface, for example resulting in direct contact with the outer surface of the cryo needle (the needle cooling surface), less cooling energy might be necessary, in particular higher temperatures and/or less application(cooling) time.

What the results also show is that adipocytes’ reaction to Staurosporine and/or the LDH assay test is not as (intense) and/or as it is known from other types of cells. This emphasizes the specific properties of adipocytes, which can in particular not be compared with tumor cells and require therefore a different treatment for initiating controlled cell death, in particular for initiation of apoptosis.

In the following to further examples are described.

Example 1 (face):

For a first example of percutaneous cryolipolysis of subcutaneous fat tissue in the face, in particular at the cheeks, according to the present invention device 10 is used, wherein first an appropriate needle assembly 11 is selected and mounted to the main body. As cryo fluid, CO2 is used and a cryo fluid source with CO2 is coupled to the main body. Change of the cryo fluid container 15 might be done. In the next step, the cryolipolysis parameters are set, wherein a needle cooling temperature (temperature of the outer cooling needle surface in the cooling area P1) of -10°C is adjusted for a cooling time of 2 minutes to initiate sufficient apoptosis A1 and necrosis A2. In particular a ratio of A1/A2 of 1 :1 is adjusted and a cooling zone CZ having a volume of 3cm³. Then the patient is prepared. After cleaning and disinfection of the skin surface, in particular around the cheek area, a local anesthetic creme with lidocaine is applied. After setup of the device, the skin is sterilized in a punctuation zone. In a next step the skin is pierced by the cryo needle 12 and the cryo needle 12 is placed with its needle cooling area under the skin in the subcutaneous tissue. Is the needle placed, the cooling process can be started by starting the cooling flow through the cryo needle 12 as adjusted. After the 2 minutes, the cryolipolysis cycle has finished and the needle 12 is removed from the tissue. At least some of the steps may be repeated and success of the treatment may be checked.

Example 2 (belly):

For a second example of percutaneous cryolipolysis according to the present invention device 10B is used, wherein first an appropriate needle assembly 11 is selected and mounted to the main body. Since a high amount of subcutaneous fat tissue at the belly of the patient has to be treated, a needle assembly comprising a needle array is selected and coupled to the main body. As cryo fluid, CO2 is used and a cryo fluid source 15 with CO2 is coupled to the main body. The cryolipolysis parameters are set, wherein a needle cooling temperature (temperature of the outer cooling needle surface in the cooling area P1) of - 40°C is adjusted for a cooling time of 3 minutes to initiate sufficient apoptosis A1 and necrosis A2. In particular a ratio of A1/A2 of 1 :1 is adjusted and a cooling zone CZ having a volume of 7,5cm³. Then the patient is prepared. After cleaning and disinfection of the skin surface, a local anesthetic creme with lidocaine is applied. In a next step the skin is pierced by the cryo needle 12 and the cryo needle 12 is placed with its needle cooling area under the skin in the subcutaneous tissue. Is the needle placed, the cooling process can be started by starting the cooling flow through the cryo needle 12 as adjusted. After 3 minutes, the cryolipolysis cycle has finished and the needle 12 is removed from the tissue. At least some of the steps may be repeated and success of the treatment may be checked. All of the features described herein with respect to one aspect of the present invention may also be realized alone or in combination in any other of the subjects according to any other aspect of the present invention, so far this is technically possible.

List of reference siqns

10, 10A, Device according to the invention for percutaneous cryolipolysis 10B, 10C 100 Kit according to the invention for percutaneous cryolipolysis

11 needle assembly

12 cryo needle

12A inner lumen for cold cryo fluid flow towards needle tip

12B inner lumen for backflow of cryo fluid

13 main body of the device

14 supply hose for cryo fluid supply and backflow

15 cryo fluid source (e.g., cryo fluid container)

16 control unit

17 operating elements for control of cryo application parameters

18 supply connector

19 adapter

20 needle hub

21 closed needle tip

22 thermal isolating material

23 lever for control of cryo fluid flow

24 cartridge filled with cryo fluid

25 end stop for limiting penetration depth

26 counter weight

27 status LED

28 display

29 temperature sensor to detect needle cooling temperature

A adipocytes (fat cells)

A1 adipocytes (fat cells) within the cooling zone died by apoptosis

A2 adipocytes (fat cells) within the cooling zone died by necrosis

CZ subcutaneous tissue cooling zone (“iceball”)

D1 outer dermis layer

D2 inner dermis layer F subcutaneous layer of body fat tissue

G cryo fluid flow

G1 cooling flow of cryo fluid

G2 backflow of cryo fluid

M body tissue (e.g. muscles)

L1..L13 possible body locations for percutaneous cryolipolysis

P1 cooling portion of cryo needle

P2 non-cooling portion of cryo needle

T1 first isothermal of cooling zone (defining boundary of cooling zone and surrounding tissue)

T2 second isothermal

T3 third isothermal

TN needle cooling temperature (cooling temperature at outer surface of cooling needle)

Claims

1 . A device (10, 10A, 10B, 10C) for percutaneous cryolipolysis, comprising: a main body (13), to which a cryo fluid source (15, 24) is couplable, and a cryo needle assembly (11 ), being coupled or couplable to the main body (13), wherein the cryo needle assembly (11) comprises at least one cryo needle (12), wherein at least one cryo needle (12) is configured to penetrate the skin (D1 , D2) of a human or an animal at least partially, wherein the device (10, 10A, 10B, 10C) is configured,

- to cause a cooling of at least one cryo needle (12) at least in a cryo needle cooling area (P1 ), when a cryo fluid source (15, 24) and the cryo needle assembly (11) are coupled to the main body (13), by using cryo fluid (G) from the cryo fluid source (15, 24), and

- to form a subcutaneous tissue cooling zone (CZ) adjacent to the cryo needle cooling area (P1) of the cooled cryo needle (12), when the cooled cryo needle (12) is penetrating the skin (D1 , D2) of a human or an animal at least partially, and wherein the device (10, 10A, 10B, 10C) is configured to cool subcutaneous fat cells (A, A1 , A2) located in the tissue cooling zone (CZ) such that at least for some of the fat cells (A, A1 , A2) located in the tissue cooling zone (CZ) apoptosis (A1) and/or necrosis (A2) is initiated.

2. The device (10, 10A, 10B, 10C) according to claim 1 , wherein the device (10, 10A, 10B, 10C) is configured to cool the at least one cryo needle (12) in the cryo needle cooling area (P1) such that for at least 5 Vol.-%, 10 Vol.-%, 15 Vol.-%, 20 Vol.-%, 25 VoL-%,30 Vol.-%, 35 Vol.-%, 40 Vol.-%, 45 VoL-%, 50 VoL-%, 55 VoL-%, 60 Vol.-%, 65 Vol.-%, 70 VoL-%, 75 VoL-%, 80 VoL-%, 85 Vol.-%, 90 Vol.-% or 95 VoL-% of the fat cells (A, A1 , A2) located in the tissue cooling zone (CZ) apoptosis (A1) is initiated.

3. The device (10, 10A, 10B, 10C) according to claim 1 , wherein the device (10, 10A, 10B, 10C) is configured to cool the at least one cryo needle (12) in the cryo needle cooling area (P1) such that for at least 5 Vol.-%, 10 Vol.-%, 15 Vol.-%, 20 Vol.-%, 25 VoL-%,30 Vol.-%, 35 Vol.-%, 40 Vol.-%, 45 VoL-%, 50 VoL-%, 55 VoL-%, 60 Vol.-%, 65 Vol.-%, 70 Vol.-%, 75 VoL-%, 80 VoL-%, 85 Vol.-%, 90 Vol.-% or 95 VoL-% of the fat cells (A, A1 , A2) located in the tissue cooling zone (CZ) necrosis (A2) is initiated.

4. The device (10, 10A, 10B, 10C) according to any one of claims 1 to 3, wherein the device (10, 10A, 10B, 10C) is configured to cool the at least one cryo needle (12) in the cryo needle cooling area (P1) such that for some fat cells located in the tissue cooling zone (CZ) apoptosis (A1 ) is initiated and for some fat cells (A, A1 , A2) located in the tissue cooling zone (CZ) necrosis (A2) is initiated, wherein in particular the ratio (A1 :A2) of the volume of fat cells (A, A1 , A2) located in the tissue cooling zone (CZ) for which apoptosis (A1 ) is initiated to the volume of fat cells (A, A1 , A2) located in the tissue cooling zone for which necrosis (A2) is initiated is in a range from 0.0, 0.1 , 0.2, 0.3, 0.4, 0.5, 0.6, 0.7. 0.8 or 0.9 to 1 .0, 0.9, 0.8, 0.7, 0.6, 0.5, 0.4, 0.3, 0.2 or 0.1.

5. The device (10, 10A, 10B, 10C) according to any one of claims 1 to 4, wherein the device (10, 10A, 10B, 10C) is configured to cool the at least one cryo needle (12) in the cryo needle cooling area (P1 ) such that in the formed tissue cooling zone (CZ) the subcutaneous tissue (F) is cold down at least to +10°C, +5°C, +4°C, +3°C, +2°C, +1 ° or 0°C, but not cold down below -50°C, -40°C, -30°C, -20°C, -10°C, -9°C, -8°C, -7°C, -6°C, -5°C, -4°C, -3°C, -2°C or -1°C, wherein the subcutaneous tissue (F) in the tissue cooling zone (CZ) is preferably cold down to a tissue temperature (T1 , T2, T3) in a range from -50°C to +10°C, -40°C to +5°C, -30°C to 0°C or -20°C to -5°C.

6. The device (10, 10A, 10B, 10C) according to any one of claims 1 to 5, wherein at least one cryo needle (12) comprises a thermal isolation (22) and/or comprises a thermal isolation material (22) or is at least partly made of a thermal isolating material (22).

7. The device (10, 10A, 10B, 10C) according to any one of claims 1 to 6, wherein the device (10, 10A, 10B, 10C) comprises a valve to control, in particular to limit, a cooling flow (G) of the cryo fluid into and/or through at least one cryo needle (12).

8. The device (10, 10A, 10B, 10C) according to any one of claims 1 to 7, wherein the device (10, 10A, 10B, 10C) is a handheld device (10, 10A, 10B, 10C).

9. The device (10, 10A, 10B, 10C) according to any one of claims 1 to 8, wherein the main body (13) is couplable to a stationary cryo fluid storage (15) as cryo fluid source (15), preferably of CO2 or N2 or N2O or a combination thereof, in particular via at least one supply line (14).

10. The device (10, 10A, 10B, 10C) according to any one of claims 1 to 9, wherein the cryo needle assembly (11) is interchangeable and detachably coupled or couplable to the main body (13), wherein in particular the main body (13) and/or the cryo needle assembly (11 ) comprises an adapter (19) via which the cryo needle assembly (11) and the main body (13) are coupled or couplable to each other.

11. The device (10, 10A, 10B, 10C) according to any one of the preceding claims, wherein at least one cryo needle (12) is a 15G, 16G, 17G, 18G, 19G, 20G, 21 G, 22G, 23G, 24G, 25G, 26G, 27G, 28G, 29G, 30G, 31 G, 32G, 33G, 34G or 35G needle or cannula.

12. The device (10, 10A, 10B, 10C) according to any one of the preceding claims, wherein the needle assembly (11 ) comprises at least two cryo needles (12), in particular a group of needles (12), in particular an array of cryo needles (12), and/or wherein the needle assembly (11 ) comprises at least one micro needle, in particular at least one micro needle array.

13. The device (10, 10A, 10B, 10C) according to any one of claims 1 to 12, wherein the device comprises a control unit (16) configured for controlling the cooling of the cryo needle cooling area (P1 ), in particular for controlling of a cooling temperature in the cryo needle cooling area (P1 ), to control the cooling temperature (T1 , T2, T3) in the tissue cooling zone (CZ).

14. A kit (100) for percutaneous cryolipolysis, comprising: a device (10, 10A, 10B, 10C), comprising: a main body (13), to which a cryo fluid source (15, 24) is couplable, and a cryo needle assembly (11 ), being coupled or couplable to the main body (13), wherein the cryo needle assembly (11) comprises at least one cryo needle (12), wherein at least one cryo needle (12) is configured to penetrate the skin (D1 , D2) of a human or an animal at least partially, wherein the device (10, 10A, 10B, 10C) is configured,

- to cause a cooling of at least one cryo needle (12) at least in a cryo needle cooling area (P1 ), when a cryo fluid source (15, 24) and the cryo needle assembly (11 ) are coupled to the main body (13), by using cryo fluid (G) from the cryo fluid source (15, 24), and

- to form a subcutaneous tissue cooling zone (CZ) adjacent to the cryo needle cooling area (P1 ) of the cooled cryo needle (12), when the cooled cryo needle (12) is penetrating the skin (D1 , D2) of a human or an animal at least partially, and wherein the device (10, 10A, 10B, 10C) is configured to cool subcutaneous fat cells (A) located in the tissue cooling zone (CZ) such that at least for some of the fat cells (A, A1 , A2) located in the tissue cooling zone (CZ) apoptosis (A1 ) and/or necrosis (A2) is initiated, and a cryo fluid source (15, 24).

15. Use of a device (10, 10A, 10B, 10C) configured for percutaneous cryolipolysis or a kit (100) configured for percutaneous cryolipolysis for cosmetic, aesthetic or therapeutic application, particularly for percutaneous cryolipolysis, the device (10, 10A, 10B, 10C) for percutaneous cryolipolysis comprising: a main body (13), to which a cryo fluid source (13, 24) is couplable, and a cryo needle assembly (11 ), being coupled or couplable to the main body (13), wherein the cryo needle assembly (11) comprises at least one cryo needle (12), wherein at least one cryo needle (12) is configured to penetrate the skin (D1 , D2) of a human or an animal at least partially, wherein the device (10, 10A, 10B, 10C) is configured,

- to form a subcutaneous (F) tissue cooling zone (CZ) adjacent to the cryo needle cooling area (P1 ) of the cooled cryo needle (12), when the cooled cryo needle (12) is penetrating the skin (D1 , D2) of a human or an animal at least partially, and wherein the device (10, 10A, 10B, 10C) is configured to cool subcutaneous (F) fat cells (A) located in the tissue cooling zone (CZ) such that at least for some of the fat cells (A, A1 , A2) located in the tissue cooling zone (CZ) apoptosis (A1) and/or necrosis (A2) is initiated.